Fusion proteins forming trimers
| Title: | Fusion proteins forming trimers |
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| Patent Number: | 8,664,366 |
| Publication Date: | March 04, 2014 |
| Appl. No: | 13/143531 |
| Application Filed: | January 09, 2009 |
| Abstract: | The present invention refers to fusion proteins comprising a neck region and carbohydrate recognition domain of a collectin trimerization domain, a linker element and an effector polypeptide. Further the invention refers to a nucleic acid encoding the said fusion protein. The fusion proteins, the nucleic acid, and the cell are suitable as pharmaceutical composition or for therapeutic, diagnostic and/or research applications as described herein. |
| Inventors: | Hill, Oliver (Neckarsteinach, DE); Branschaedel, Marcus (Laupheim, DE); Gieffers, Christian (Dossenheim, DE); Thiemann, Meinolf (Schriesheim, DE) |
| Assignees: | Apogenix GmbH (Heidelberg, DE) |
| Claim: | 1. A fusion protein comprising (i) a collectin family trimerization domain consisting of amino acid residues 221-375 of SEQ ID NO: 21, optionally further consisting of an amino acid substitution at residue 355; (ii) a linker element; and (iii) an effector polypeptide selected from the group consisting of a single chain antibody, an antigen binding fragment of a single chain antibody, or an antigen binding fragment of an antibody, wherein the effector polypeptide is located N-terminally of the collectin family neck region wherein said linker element has 25 or less amino acids. |
| Claim: | 2. The fusion protein claim 1 , wherein the collectin family trimerization domain consists of the amino acid residues 221-375 of SEQ ID NO: 21. |
| Claim: | 3. The fusion protein of claim 1 , wherein the amino acid residue 355 in SEQ ID NO: 21 is optionally substituted to a polar amino acid selected from the group consisting of aspartic acid, glutamic acid, asparagine and glutamine. |
| Claim: | 4. The fusion protein of claim 1 , wherein the effector polypeptide is a single chain antibody against IL4R-alpha. |
| Claim: | 5. The fusion protein of claim 1 , wherein the linker element has the amino acid sequence of SEQ ID NO: 56, which is GSSGSSGSSGSG. |
| Claim: | 6. The fusion protein of claim 1 , wherein the linker element has the amino acid sequence of SEQ ID NO: 34, which is GSSGSSGSSGS. |
| Claim: | 7. The fusion protein of claim 1 , further comprising an N-terminal signal peptide domain. |
| Claim: | 8. The fusion protein of claim 1 , further comprising a recognition/purification domain selected from the group consisting of a strep-tag domain and a poly-His domain. |
| Claim: | 9. The fusion protein of claim 1 , wherein the effector polypeptide is a monovalent Fab fragment. |
| Claim: | 10. The fusion protein of claim 1 , wherein the effector polypeptide is a single chain variable fragment (scFv). |
| Claim: | 11. The fusion protein of claim 10 , wherein the scFv has a variable domain of a heavy chain located N-terminal to the a variable domain of a light chain. |
| Claim: | 12. A trimeric complex consisting of three identical fusion proteins of claim 1 . |
| Current U.S. Class: | 5303/873 |
| Patent References Cited: | 2004/0047873 March 2004 Al- Shamkhani et al. 2004/0197876 October 2004 Tschopp et al. 2004/0247563 December 2004 Lynch et al. 2009/0325867 December 2009 Cohen et al. 2010/0199364 August 2010 Hill et al. 2010/0322922 December 2010 Martin-Villalba et al. 2011/0111494 May 2011 Hill et al. WO 95/31540 November 1995 WO 97/01633 January 1997 WO 01/42298 June 2001 WO 02/090553 November 2002 WO 03/086301 October 2003 WO 2004/024925 March 2004 WO 2007/102690 September 2007 WO 2009/007120 January 2009 |
| Other References: | Crouch et al., (JBC 2006, 281:18008-18014). cited by examiner Hakansson, K. and K.B. Reid (2000). “Collectin structure: a review [In Process Citations]”,Protein Science 9: pp. 1607-1617. cited by applicant Haswell, et al. (2001). “Analysis of the oligomeric requirement for signaling by CD40 using soluble multimeric forms of its ligand, CD154”, Eur. J. Immunol. 31: 3094-3100. cited by applicant Sano, H. and Y. Kuroki (2005). “The lung colectins, SP-A and SP-D, modulate pulmonary innate immunity”, 42: pp. 279-287. cited by applicant Kishore, et al. (2006). “Surfactant proteins SP-A and SP-D: Structure, function and receptors”, Molecular Immunology, 43: pp. 1293-1315. cited by applicant Wu et al.: Molecular Immunology 46:2381-2388, 2009. cited by applicant Holler, N. et al.: “Two Adjacent Trimeric Fas Ligands Are Required for Fas Signaling and Formation of a Death-Inducing Signaling Complex”, Molecular and Cellular Biology, American Societv for Microbiology. vol. 23 No. 4 Feb. 1, 2003, p. 1428-1440. cited by applicant Hoppe, H.-J. et al.: “A parallel three stranded a-helical bundle at the nucleation site of collagen triple-helix formation”, FEBS Letters, vol. 344, No. 2/03, Jan. 1, 1994, pp. 191-195. cited by applicant Kornbluth, R. S. et al.: “CD40L (CD154) fusion protein with pulmonary surfactant protein D as a prototype for soluble multimeric TNF superfamily ligand molecules”, FASEB Journal, Fed. of American Soc. for Experimental Biology. vol. 14 No. 6, Apr. 20, 2000. cited by applicant |
| Primary Examiner: | Stanfield, Cherie M |
| Attorney, Agent or Firm: | Perkins Coie LLP Kung, Viola T. |
| Accession Number: | edspgr.08664366 |
| Database: | USPTO Patent Grants |
| Language: | English |
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